A reliable qPCR technique for detecting viable Xanthomonas arboricola pv. pruni cells

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Date

2024

Authors

Sabuquillo, Pilar
Berruete Rodríguez, Isabel María
Cubero, Jaime
Palacio Bielsa, Ana

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Abstract

Xanthomonas arboricola pv. pruni (Xap) is the causal agent of bacterial spot of stone fruits and almond (Prunus spp). Detection of Xap is typically carried out using quantitative real-time PCR (qPCR) combined with culture-based isolation. However, qPCR does not differentiate between viable and dead cells, potentially leading to an overestimation of the infective population in a sample. Such overestimation could result in unnecessary phytosanitary measures. The present study aims to develop a specific protocol ideally targeting to detection of only live Xap bacterial cells. To address this challenge, the viable quantitative PCR (v-qPCR) method was evaluated using three nucleic acid-binding dyes: propidium monoazide (PMA), a combination of PMA and ethidium monoazide (EMA), and PMAxx™, an improved version of PMA. PMAxx™ proved to be the most suitable dye for the detection and quantification of living bacterial cells. This methodology was also evaluated in infected plant material over time and can be considered a rapid and reliable alternative to PCR methods for detecting only those putative infective Xap that may pose a risk for Prunus crops.

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Bibliographic citation

Sabuquillo, P., Berruete, I. M., Cubero, J., & Palacio-Bielsa, A. (2024). A reliable qPCR technique for detecting viable Xanthomonas arboricola pv. Pruni cells. Applied Microbiology and Biotechnology, 108(1), 472. https://doi.org/10.1007/s00253-024-13288-y
AGROVOC subjects
Xanthomonas arboricola pv. pruni
Prunus
Frutas de hueso
Árboles frutales
Enfermedades bacterianas
Colorantes
Reacción en cadena de polimerasa cuantitativa
Protocolos

Other field subjects
Árboles frutales
Colorantes
Enfermedades bacterianas
Frutas De Hueso
Protocolos
Prunus
Reacción en cadena de polimerasa cuantitativa
Xanthomonas arboricola pv. pruni

Sponsorship

Este trabajo fue financiado por el proyecto PID2021-123600ORC44, con el apoyo de MICIU/AEI/https://doi.org/10.13039/501100011033 y por el FEDER, UE

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http://hdl.handle.net/10532/7244

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